The ability of a particular antibody to combine with one antigen instead of another.
specificity
fit between the antigenic determinant and the combining site of the antibody.
non-covalent force
some of the determinants of an antigen are shared by apparently unrelated molecules.
cross-reactivity
strength of a multivalent antibody to bind to a multivalent antigen
avidity
the bond between a single antigenic determinant and an individual combining site of an antibody.
affinity
excessive amount of antibody that can give a false negative result.
prozine
decreased amount of antibodyt caused by an excess of antigen
postzone
point of equal antigen and antibody amounts which yield the greatest reaction.
zone of equivalence
deviance from the optimum pH of ___ will alter reactivity of the antibody.
7.0
different classes of antibodies react at different __________.
temperatures
what is the antibody that reacts at warm temperatures?
IGG
what is the antibody that reacts at cold temperatures?
IgM
incubatin time varies with the _____ of antibody.
class
which types of antibodies will be all the same class and have only one type of heavy and light chain?
monoclonal antibodies
these are uniform, highly specific antibodies that caon be produced in large quantitites, cloned from a single cell to bind to a single specific antigen.
Monoclonal antibodies
what are the three uses of monoclonal antibodies?
typing erythrocyte antigens
typing leukocyte antigens
as a reagent
serological samples should be centrifuged for __ minutes at moderate speed.
10
if testing cannot be done promptly the serum should be frozen at what temperature?
20oC
how is complement inactivated?
heat serum to 56oC for 30min
if serum is not used within four hours, reheat serum at 56oC for ___ minutes.
10
the highest dilution in which a reaction occurs.
titer
what is the purpose of a titer?
to determine the concentration of antibody
this is used to make serum less concentrated.
diluent
each succeeding dilution is the same as the preceding dilution
fold
calculations of any given dilution can be determined by using this formula:
D1(V1)=D2(V2)
in the formula D1(V1)=D2(V2) what is the starting dilution.
D1
in the formula D1(V1)=D2(V2) what is the starting volume?
V1
in the formula D1(V1)=D2(V2) what is the final dilution
D2
in the formula D1(V1)=D2(V2) what is the final total volume?
V2
pipettes should be periodically calibrated for what three things?
accuracy
precision
reproducibility
Red Blood cell Suspensions should be used in serological tests as an ________.
indicator
in an RBC suspension what should be used as the diluent?
2-5% NaCl
RBC's contain _______; serve as receptor sites for antibodies or complement.
antigens
what are the four carriers used in serology?
carbon
RBC's
Latex
Bacteria
what is the calucation that can be used to determine the minimum packed cell volume needed to make a solution?
PCV= (total volume X % desired)/100
in an RBC suspension this represents the packed RBCs.
PCV
true or false
Hemolyzed RBC's can be used in an RBC suspension.
False
Mono and ASO use what carrier?
Latex
Floculation tests use what carrier?
Charcoal
what carrier is used in hemaglutination?
RBC's
in what testing method, if antigens are present in serum, they will bind with antibodies forming visible cross-linked aggregates.
latex agglutination
in latex agglutination antibody molecules bind to latex _____.
beads
what are the five latex agglutination immunologic assays?
antistreptolysin O
Mono Test
C-reactive protein
IgG and IgM rheumatoid factors
Rubella antibody
in what test are latex particles coated with strptolysin O antigen?
Antistreptolysin O
in Antistreptolysin O visible agglutination will be seen mixed with patient's serum containing what?
ASO antibody
titers begin to rise at __ days after the onset of infection and peaks at ____ weeks after infection.
7 days
3-6 weeks
In ASO, a four fold increase between acute and convalescent tube indicates a recent infection with what?
Group A strep
ASO antibodies are elevated in acute/convalescent sera with what disease?
acute rheumatic fever
serological testing should compare acute and convalescent sera collected ______ weeks apart.
three
how long are ASO samples stable at 2-8oC?
eight days
how long are ASO specimens stable at -25oC?
3 months
this is based on agglutination of horse RBC's by hetrophil antibody present in infectious mononucleosis.
Monotest
in monotest horse RBC's exhibit antigens against both __________ and ______________ threfore a differential absorption of patient serum is necessary.
forssman
infectious mononucleosis
the monotest immunologic theory is based on __________ principle.
Davidshon principle
in the monotest immunologic theory, what contains only forssman antigen which will absorb forssman heterophil antibody?
guinea pig kidney
in monotest immunologic theory what will absorb only no-forssman heterophil antibody of infectious mononucleosis?
beef RBC's
in a monotest immunologic theory this is tested with both the guniea pig-serum specimen and the beef RBC-serum specimen.
horse RBC's
to be positive for IM heterophil antibody, the guinea pig should be _________ for agglutination and the beef RBC should be _________ for agglutination.
Guinea pig: positive
Beef RBC: negative
what is the sample for Monotest?
Serum or plasma mixed with EDTA
what is the confirmatory test for infectious mononucleosis?
Epstein-Barr virus antibody
this is the causitive agent of infectious mononucleosis?
ebstein-barr virus
this is a DNA herpes-type virus that infects the B-lymphocytes?
EBV
this is tested to confirm infectious mononucleosis on immune suppressed patients.
EBV antibodies
this test is beneficial in defining immune status and stage of infectious mononucleosis.
EBVA
antigen-antibody __________ when combined in proportions at or near equivalence.
precipitate
in what type of test is interaction of soluble antigen with antibody that results in the formation of a precipitate of fine particles?
flocculation
in flocculation test particles are ____________ or _____________ visible.
macroscopically
microscopically
what are two examples of flocculation tests?
VDRL
RPR
this test is based on agglutination of RBC's with antigens that are natural or bound to RBC's
hemagglutination
what are three examples of hemagglutination immunologic assays?
ABO antibodies
Rh antigens
Cold Agglutinins
Precipitation methods are performed in what two mediums?
semi solid media (agar or agarose)
non-gel support medium (cellulose acetate)
what are the two types of precipitation methods?
double immunodifusion
radial immunodifusion
in this test antibody dilutions and specific antigens are placed in adjacent wells. Antigen and antibody diffuse out, bind together to form a visible precipitate.
Double immunodiffusion
what are the three basic reaction patterns in double immunodiffusion?
Identity
Non-identity
partial identity
In which double immunodiffusion result in precipitin band forms a single smooth arch?
Identity
in which double immunodiffusion resuld do precipitin lines merge in a spur formation?
partial identity
in which double immunodiffusion reslut do precipitin lines cross each other?
non-identity
what are the acceptable specimens for double immunodiffusion?
serum
urine
in double immunodiffusion why are urine specimens tested in concentrated and unconcentrated forms?
wide range of light cahin concentrations
what are the two double immunodiffusion immunologic assays?
Rheumatoid arthritis
systemic lupus erythrematosus
Radial immunodiffusion (RID) is used to identify and quantitate the number of _________ found in human serum.
proteins
in radial immuno diffusion (RID) the antigen will diffuse throughout the agan and form what?
a precipitin ring
in radial immunodiffusion specific antibody is added to a buffered agarose medium, test ________ is added to a well in the agarose.
antigen
in radial immunodiffusion (RID) diameter of the precipitin rig is related to what?
antigen concentration
what are the four clinical applications of radial immunodiffusion (RID)?
Alpha-1-antitrypsin
Transferrin
C3
Immunoglobulin
what are the two electro immunodiffusion methods?
Countercurrent immunoelectrophoresis (CIE)
rocket immunoelectrophoresis
in what test do antigens and antibodies move more quickly toward each other with addition of electrical current?
countercurrent immunoelectrophoresis (CIE)
in CIE agarose gel is used. pH is such that antibodies are ________ charged and antigens are ________ charged.
antibodies: positively
antigen: negatively
in countercurrent immiunoelectrophoresis how is the zone of equivalence represented?
antigen-antibody complex forsm a visible line of precipitin
what is the clinical application of countercurrent immunoelectrophoresis (CIE)?
detection of anti-nuclear ribonucleoprotein (nRNP):
associated to sharp's syndrome (mixed connective tissue disease)
what test is associated with sharp's syndrome (mixed conective tissue disease)?
detection of anti-nuclear ribonucleoprotein (nRNP)
in rocket electrophoresis is based on antigen ______________ in an antibody containing gel.
electrophoresis
in rocket electrophoresis what is adjusted to inhibit antibody migration?
pH
in rocket electrophoresis the height of the rocket is __________ to the antigen concentration.
proportional
what ar the four types of labeling techniques?
radio immuno assay (RIA)
chemiluminescence
enzyme immunoassay (EIA)
immunofluorescent techniques
what are the two assays that involve chemiluminescence?
competitive assay
sandwich assay
what are the two types of immunofluorescent techniques?
direct
indirect
in what test are radioisotopes used to measure the concentration of antigens or antibodies
radioimmunoassay (RIA)
in what test are radiolabled ligands bound to antigen-antibody complex?
radioimmunoassay
in RIA radiactivity is counted on what?
gamma counter
this test is extremly sensitive and able to detect trace amounts of antigen or antibody.
radioimmunoassay (RIA)
what are the two clinical applications of radioimmunoassay (RIA)
hepatitis A IgM antibody
hepatitis A antigen
in this test lables can be attached to antigen or antibody.
chemiluminescence
in this chemiluminescence assay, a fixed amount of labeled antigen competes with unlabeled antigen (patient sample) for a limited numberr of antibody-binding sites.
competitive assay
in a competitive assay the amount of light emitted is _________ proportional to the amount of analyte (antigen) being tested.
inversely
in this chemiluminescene test sample antigen binds to a fixed antibody on a solid phase, then a second antibody labled with chemiluminescent binds to the antigen-Ab complex on the solid phase.
sandwich assay
in a sadwich assay the amount of light emitted is __________ proportional to the amount of the analyte (antigen) being tested.
directly
what are the two clinical applications of chemiluminescence?
hepatitis A IgM antibody
hepatitis A antigen
this test is similar to RIA but used a non-isotopic label.
enzyme immunoassay
this test is safer than RIA yet has the same specificity and sensitivity as RIA.
enzyme immunoassay (EIA)
in Enzyme immunoassay (EIA) what is used instead of a radioisotope?
an enzyme labeled antibody or antigen conjugate
what ar the most commonly used enzymes to detect the presence and quantity of antigen or antibody in patient serum?
peroxidase
alkaline phosphatase
in EIA substrate causes color production and is measured how?
spectrophotometry
what are the three clinical applications of Enzyme Immunoassay (EIA)?
hepatitis B
HIV antibody
Cytomegalovirus antigen/antibody
in this extremely specific and sensitive test, fluorescent molecules are used instead of radioisotopes or enzyme labels.
immunofluorescent techniques
what are the three clinical applications of immunofluorescent techniques?
CMV antigen
CMV antibodies
HIV antibodies
in this immunofluorescene technique conjugated reagent antibody is used to detect antigen-antibody complex at microscope level.
Direct Immunofluorescence
in direct immunofluorescence what is used ot visualized many bacteria in direct smears?
fuorescein-conjugated antibodies boud to fuorochrome
direct immunofluorescence uses the immunological/physiological theory of:
intracellular antigen detection
what are the two sepcimens used for direct immunofluorescence?
tissue
bacteria smears
in this test, antigen source such as toxoplasma microorganism or virus of infected tissue culture cells to the specific antibody being tested is affixed to a slide.
indirect immunofluorescence
in indirect immunofluorescence what reacts with antigen forming an antigen-antibody complex?
unkown antibody (patient's Ab)
in indirect immunofluorescence what is added to the Ab-Ag complex to bind to the antibody as a marker?
labeled anti-immunoglobulin (fluorescent Ab)
indirect immunofluorescence measures what?
antibody
direct immunofluorescene measures what?
antigen
what is the specimen collected for indirect immunofluorescence?
serum
what are three indications for performing indirect immunofluorescene on serum?
toxoplasma microorganism
virus infected tissue culture cells
ANA antibodies
what is the classic method for demonstrating presence of a complement fixing antibody in serum?
complement fixation
what is the indicator system for complement fixation?
sheep RBC's
anti-sheep antibody
complement from guinea pig serum
in complement fixation patient serum is added to _______ antigen, and complement is added to the solution.
known
in complement fixation, if serum contains the antibody for the antigen, then an antigen-antibody complex will bind with what?
complement
in complement fixation sheep RBC's with anti-sheep antibody are added to the solution of antibodies, antigen, and complement. if complement has not been bound, it is available to bind with what?
coated sheep RBC's
hemolysis of indicator sheep cells indicates a lack of antibody in patient serum and a _________ complement fixation test.
negative
how is a positive complement fixation test demonstrated.
lack of hemolysis
why is a positive complement fixation test demonstrated by the lack of hemolysis in indicator sheeps RBC's?
patients serum does contain a complement fixing antibody
what are the three clinical applications of complement fixation?
Varicella-zoster virus
Rubella virus
Herpes simplex infection
this is an in vitro method that amplifies low level of specific DNA sequences in a sample to higher quantities.
polymerase Chain Reaction
in PCR ___ short DNA primers are used as templates that follow the enzymatic process.
two
in PCR the enzymatic prcess is carried out in cycles and repeated up to ___ times.
30
what are the four steps of enzymatic process in PCR?
DNA denaturing
Primer anealing
Extension of primed DNA sequenced
The amount specific DNA sequence double and examined in various ways
polymerase chain reaction is used in the detection of ____.
DNA
what are the two clinical applications for PCR?
human papilloman virus (HPV)
coronary artery disease
If the nucleic acid of interest is RNA rather than DNA, the PCR procedures can be modified to include the conversion of RNA to DNA using what?
Reverse Transcriptase-Polymerase (RT-PCR)
the clinical application for RT-PCR is to test for what?